The blood-brain barrier (BBB) is a specialized structure that controls the movement of substances between the bloodstream and the brain. Investigating BBB permeability is crucial for developing effective treatments for various central nervous system (CNS) disorders. In vitro BBB models offer several advantages over animal models, including high-throughput analysis, accessibility, and ethical considerations. Spheroid-based models have been proposed as an in vitro model for the BBB, but they require a specialized culture medium containing growth factors, making them cost-prohibitive. In this study, a novel method for constructing spheroid-based BBB models (BBBoids) is presented that does not require a specialized medium or growth factors, increasing affordability and cost-effectiveness. BBBoids comprise primary murine astrocytes which form the spheroid core, and the bEnd.3 endothelial cell line, which encases the spheroid surface, acting as a barrier. BBBoids form within 6 days and exhibit barrier properties and proper morphology for up to 2 weeks. This model is suitable for investigating the effects of various substances on BBB integrity and testing the permeability of different therapeutic agents. The BBBoid platform is a reliable, adaptable, and economical in vitro instrument that may expedite the study of potential therapeutics for a range of neuropathologies.
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